title: ATF2 knockdown reinforces oxidative stress-induced apoptosis in TE7 cancer cells.
abstract: Cancer cells showing low apoptotic effects following oxidative stress-induced DNA damage are mainly affected by growth arrest. Thus, recent studies focus on improving anti-cancer therapies by increasing apoptosis sensitivity. We aimed at identifying a universal molecule as potential target to enhance oxidative stress-based anti-cancer therapy through a switch from cell cycle arrest to apoptosis. A cDNA microarray was performed with hydrogen peroxide-treated oesophageal squamous epithelial cancer cells TE7. This cell line showed checkpoint activation via p21(WAF1) , but low apoptotic response following DNA damage. The potential target molecule was chosen depended on the following demands: it should regulate DNA damage response, cell cycle and apoptosis. As the transcription factor ATF2 is implicated in all these processes, we focused on this protein. We investigated checkpoint activation via ATF2. Indeed, ATF2 knockdown revealed ATF2-triggered p21(WAF1) protein expression, suggesting p21(WAF1) transactivation through ATF2. Using chromatin immunoprecipitation (ChIP), we identified a hitherto unknown ATF2-binding sequence in the p21(WAF1) promoter. p-ATF2 was found to interact with p-c-Jun, creating the AP-1 complex. Moreover, ATF2 knockdown led to c-Jun downregulation. This suggests ATF2-driven induction of c-Jun expression, thereby enhancing ATF2 transcriptional activity via c-Jun-ATF2 heterodimerization. Notably, downregulation of ATF2 caused a switch from cell cycle arrest to reinforced apoptosis, presumably via p21(WAF1) downregulation, confirming the importance of ATF2 in the establishment of cell cycle arrest. 1-Chloro-2,4-dinitrobenzene also led to ATF2-dependent G2/M arrest, suggesting that this is a general feature induced by oxidative stress. As ATF2 knockdown also increased apoptosis, we propose ATF2 as a target for combined oxidative stress-based anti-cancer therapies.
authors.affiliation: Department of Pathology, Otto-von-Guericke University, Magdeburg, Germany.
authors.forename: Diana
authors.initials: D
authors.lastname: Walluscheck
authors.forename: Angela
authors.initials: A
authors.lastname: Poehlmann
authors.forename: Roland
authors.initials: R
authors.lastname: Hartig
authors.forename: Uwe
authors.initials: U
authors.lastname: Lendeckel
authors.forename: Peter
authors.initials: P
authors.lastname: Schönfeld
authors.forename: Agnes
authors.initials: A
authors.lastname: Hotz-Wagenblatt
authors.forename: Kathrin
authors.initials: K
authors.lastname: Reissig
authors.forename: Khuloud
authors.initials: K
authors.lastname: Bajbouj
authors.forename: Albert
authors.initials: A
authors.lastname: Roessner
authors.forename: Regine
authors.initials: R
authors.lastname: Schneider-Stock
mesh_terms: D051698:Activating Transcription Factor 2 / Q000378:metabolism*; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D001665:Binding Sites; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D045744:Cell Line, Tumor; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000378:metabolism; D004249:DNA Damage / Q000235:genetics; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D055785:Gene Knockdown Techniques; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008954:Models, Biological; D018384:Oxidative Stress / Q000187:drug effects / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding / Q000187:drug effects; D016755:Proto-Oncogene Proteins c-jun / Q000378:metabolism
chemical_list: C497105:ATF2 protein, human; D051698:Activating Transcription Factor 2; D050759:Cyclin-Dependent Kinase Inhibitor p21; D016755:Proto-Oncogene Proteins c-jun; D006861:Hydrogen Peroxide
keywords: ATF2 knockdown; cell cycle arrest; combined treatment; increase in apoptosis sensitivities; oxidative stress-induced DNA damage; p21WAF1